Quantitative Hemolysis Assays

Methods Mol Biol. 2021:2341:25-30. doi: 10.1007/978-1-0716-1550-8_4.

Abstract

Many strains of Staphylococcus aureus produce a variety of cytolysins that target many different cell types to both fight the immune system and acquire nutrients. This includes hemolysins which destroy erythrocytes and are well studied virulence factors. Traditionally, hemolysin activity is measured on blood agar plates due to the simplicity of the assay. While this is telling, it cannot encapsulate the full story because S. aureus is known to behave differently in broth and on agar. Furthermore, plate-based assays are primarily semiquantitative and often a more accurate determination of hemolytic potential is needed to discern differences between strains. Here, we describe a method to quantify hemolysin activity from broth or similarly grown cells.

Keywords: Alpha-hemolysin; Hemolysin; Red blood cell lysis; Staphylococcus aureus; Toxin.

MeSH terms

  • Animals
  • Bacterial Proteins / analysis
  • Bacterial Proteins / metabolism
  • Culture Media / chemistry
  • Erythrocytes / physiology*
  • Hemolysin Proteins / analysis*
  • Hemolysin Proteins / metabolism
  • Hemolysis
  • Humans
  • Staphylococcus aureus / growth & development*
  • Staphylococcus aureus / metabolism
  • Staphylococcus aureus / pathogenicity
  • Virulence Factors / analysis
  • Virulence Factors / metabolism

Substances

  • Bacterial Proteins
  • Culture Media
  • Hemolysin Proteins
  • Virulence Factors